Protozoa: Structure, Classification, Growth, and Development

Robert G. Yaeger

General Concepts
Protozoa Protozoa are one-celled animals found worldwide in most habitats. Most species are free living, but all higher animals are infected with one or more species of protozoa. Infections range from asymptomatic to life threatening, depending on the species and strain of the parasite and the resistance of the host. Structure Protozoa are microscopic unicellular eukaryotes that have a relatively complex internal structure and carry out complex metabolic activities. Some protozoa have structures for propulsion or other types of movement. Classification On the basis of light and electron microscopic morphology, the protozoa are currently classified into six phyla. Most species causing human disease are members of the phyla Sacromastigophora and picomplexa. ife Cycle Sta!es !he stages of parasitic protozoa that actively feed and multiply are fre"uently called trophozoites# in some protozoa, other terms are used for these stages " Cysts are stages with a protective membrane or thic$ened wall. Protozoan cysts that must survive outside the host usually have more resistant walls than cysts that form in tissues. #eproduction %inary fission, the most common form of reproduction, is asexual# multiple asexual division occurs in some forms. %oth sexual and asexual reproduction occur in the picomplexa. $utrition ll parasitic protozoa re"uire preformed organic substancesthat is, nutrition is holozoic as in higher animals.

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%$&#'D(C&%'$ !he Protozoa are considered to be a sub$ingdom of the $ingdom Protista, although in the classical system they were placed in the $ingdom nimalia. More than )*,*** species have been described, most of which are free'living organisms# protozoa are found in almost every possible habitat. !he fossil record in the form of shells in sedimentary roc$s shows that protozoa were present in the Pre'cambrian era. nton van (eeuwenhoe$ was the first person to see protozoa, using microscopes he constructed with simple lenses. %etween &)*+ and &*&), he described, in addition to free'living protozoa, several parasitic species from animals, and Giardia lamblia from his own stools. ,irtually all humans have protozoa living in or on their body at some time, and many persons are infected with one or more species throughout their life. Some species are considered commensals, i.e., normally not harmful, whereas others are patho!ens and usually produce disease. Protozoan diseases range from very mild to life'threatening. Individuals whose defenses are able to control but not eliminate a parasitic infection become carriers and constitute a source of infection for others. In geographic areas of high prevalence, well'tolerated infections are often not treated to eradicate the parasite because eradication would lower the individual-s immunity to the parasite and result in a high li$elihood of reinfection. Many protozoan infections that are inapparent or mild in normal individuals can be life'threatening in immunosuppressed patients, particularly patients with ac"uired immune deficiency syndrome . I/S0. 1vidence suggests that many healthy persons harbor low numbers of Pneumocystis carinii in their lungs. 2owever, this parasite produces a fre"uently fatal pneumonia in immunosuppressed patients such as those with I/S. Toxoplasma gondii, a very common protozoan parasite, usually causes a rather mild initial illness followed by a long'lasting latent infection. I/S patients, however, can develop fatal toxoplasmic encephalitis. Cryptosporidium was described in the &3th century, but widespread human infection has only recently been recognized. Cryptosporidium is another protozoan that can produce serious complications in patients with I/S. Microsporidiosis in humans was reported in only a few instances prior to the appearance of I/S. It has now become a more common infection in I/S patients. s more thorough studies of patients with I/S are made, it is li$ely that other rare or unusual protozoan infections will be diagnosed. Acanthamoeba species are free'living amebas that inhabit soil and water. 4yst stages can be airborne. Serious eye'threatening corneal ulcers due to Acanthamoeba species are being reported in individuals who use contact lenses. !he parasites presumably are transmitted in contaminated lens' cleaning solution. mebas of the genus Naegleria, which inhabit bodies of fresh water, are responsible for almost all cases of the usually fatal disease

primary amebic meningoencephalitis. !he amebas are thought to enter the body from water that is splashed onto the upper nasal tract during swimming or diving. 2uman infections of this type were predicted before they were recognized and reported, based on laboratory studies of canthamoeba infections in cell cultures and in animals. !he lac$ of effective vaccines, the paucity of reliable drugs, and other problems, including difficulties of vector control, prompted the 6orld 2ealth Organization to target six diseases for increased research and training. !hree of these were protozoan infectionsmalaria, trypanosomiasis, and leishmaniasis. lthough new information on these diseases has been gained, most of the problems with control persist. Structure Most parasitic protozoa in humans are less than )* +m in size. !he smallest .mainly intracellular forms0 are & to &7 8m long, but Balantidium coli may measure &97 8m. Protozoa are unicellular eu$aryotes. s in all eu$aryotes, the nucleus is enclosed in a membrane. In protozoa other than ciliates, the nucleus is vesicular, with scattered chromatin giving a diffuse appearance to the nucleus, all nuclei in the individual organism appear ali$e. One type of vesicular nucleus contains a more or less central body, called an endosome or karyosome. !he endosome lac$s /: in the parasitic amebas and trypanosomes. In the phylum picomplexa, on the other hand, the vesicular nucleus has one or more nucleoli that contain /: . !he ciliates have both a micronucleus and macronucleus, which appear "uite homogeneous in composition. !he organelles of protozoa have functions similar to the organs of higher animals. !he plasma membrane enclosing the cytoplasm also covers the pro,ectin! locomotory structures such as pseudopodia, cilia, and fla!ella. !he outer surface layer of some protozoa, termed a pellicle, is sufficiently rigid to maintain a distinctive shape, as in the trypanosomes and Giardia. 2owever, these organisms can readily twist and bend when moving through their environment. In most protozoa the cytoplasm is differentiated into ectoplasm .the outer, transparent layer0 and endoplasm .the inner layer containing organelles0# the structure of the cytoplasm is most easily seen in species with pro;ecting pseudopodia, such as the amebas. Some protozoa have a cytosome or cell <mouth< for ingesting fluids or solid particles. 4ontractile vacuoles for osmoregulation occur in some, such as Naegleria and Balantidium. Many protozoa have subpellicular microtubules# in the picomplexa, which have no external organelles for locomotion, these provide a means for slow movement. !he trichomonads and trypanosomes have a distinctive undulating membrane between the body wall and a flagellum. Many other structures occur in parasitic protozoa, including the =olgi apparatus, mitochondria, lysosomes, food vacuoles, conoids in the picomplexa, and other specialized structures. 1lectron microscopy is essential to visualize the details of protozoal structure. >rom the point of view of functional and physiologic complexity, a protozoan is more li$e an animal than li$e a single

cell. >igure **'& shows the structure of the bloodstream form of a trypanosome, as determined by electron microscopy.

-%G(#. //-0 -ine structure of a protozoan parasite, Trypanosoma evansi, as revealed 1y transmission electron microcopy of thin sections" . dapted from ,ic$erman @A Protozoology. ,ol. ? (ondon School of 2ygiene and !ropical Medicine, (ondon, &3**, with permission.0 Classification In &3B9 the Society of Protozoologists published a taxonomic scheme that distributed the Protozoa into six phyla. !wo of these phylathe

ife Cycle Sta!es

/uring its life cycle, a protozoan generally passes through several stages that differ in structure and activity. &rophozoite .=ree$ for <animal that feeds<0 is a general term for the active, feeding, multiplying stage of most protozoa. In parasitic species this is the stage usually associated with pathogenesis. In the hemoflagellates the terms amastigote, promastigote, epimastigote, and trypomastigote designate trophozoite stages that differ in the absence or presence of a flagellum and in the position of the $inetoplast associated with the flagellum. variety of terms are employed for stages in the picomplexa, such as tachyzoite and bradyzoite for Toxoplasma gondii. Other stages in the complex asexual and sexual life cycles seen in this phylum are the merozoite .the form resulting from fission of a multinucleate schizont0 and sexual stages such as !ametocytes and !ametes. Some protozoa form cysts that contain one or more infective forms. Multiplication occurs in the cysts of some species so that excystation releases more than one organism. >or example, when the trophozoite of Entamoeba histolytica first forms a cyst, it has a single nucleus. s the cyst matures nuclear division produces four nuclei and during excystation four uninucleate metacystic amebas appear. Similarly, a freshly encysted Giardia lamblia has the same number of internal structures .organelles0 as the trophozoite. 2owever, as the cyst matures the organelles double and two trophozoites are formed. 4ysts passed in stools have a protective wall, enabling the parasite to survive in the outside environment for a period ranging from days to a year, depending on the species and environmental conditions. 4ysts formed in tissues do not usually have a heavy protective wall and rely upon carnivorism for transmission. Oocysts are stages resulting from sexual reproduction in the picomplexa. Some apicomplexan oocysts are passed in the feces of the host, but the oocysts of Plasmodium, the agent of malaria, develop in the body cavity of the mos"uito vector.

#eproduction Ceproduction in the Protozoa may be asexual, as in the amebas and flagellates that infect humans, or 1oth asexual and sexual, as in the picomplexa of medical importance. !he most common type of asexual multiplication is 1inary fission, in which the organelles are duplicated and the protozoan then divides into two complete organisms. Division is lon!itudinal in the fla!ellates and transverse in the ciliates2 amebas have no apparent anterior'posterior axis. .ndodyo!eny is a form of asexual division seen in Toxoplasma and some related organisms. !wo daughter cells form within the parent cell, which then ruptures, releasing the smaller progeny which grow to full size before repeating the process. In

schizo!ony, a common form of asexual division in the picomplexa, the nucleus divides a number of times, and then the cytoplasm divides into smaller uninucleate merozoites" In Plasmodium, Toxoplasma, and other apicomplexans, the sexual cycle involves the production of gametes .!amo!ony0, fertilization to form the zygote, encystation of the zygote to form an oocyst, and the formation of infective sporozoites .sporogony0 within the oocyst.

Some protozoa have complex life cycles re"uiring two different host species# others re"uire only a single host to complete the life cycle. single infective protozoan entering a susceptible host has the potential to produce an immense population. 2owever, reproduction is limited by events such as death of the host or by the host-s defense mechanisms, which may either eliminate the parasite or balance parasite reproduction to yield a chronic infection. >or example, malaria can result when only a few sporozoites of Plasmodium falciparumperhaps ten or fewer in rare instancesare introduced by a feeding Anopheles mos"uito into a person with no immunity. Cepeated cycles of schizogony in the bloodstream can result in the infection of &7 percent or more of the erythrocytesabout +77 million parasites per milliliter of blood. $utrition !he nutrition of all protozoa is holozoic# that is, they re"uire organic materials, which may be particulate or in solution. mebas engulf particulate food or droplets through a sort of temporary mouth, perform digestion and absorption in a food vacuole, and e;ect the waste substances. Many protozoa have a permanent mouth, the cytosome or micropore, through which ingested food passes to become enclosed in food vacuoles. Pinocytosis is a method of ingesting nutrient materials whereby fluid is drawn through small, temporary openings in the body wall. !he ingested material becomes enclosed within a membrane to form a food vacuole. Protozoa have metabolic pathways similar to those of higher animals and re"uire the same types of organic and inorganic compounds. In recent years, significant advances have been made in devising chemically defined media for the in vitro cultivation of parasitic protozoa. !he resulting organisms are free of various substances that are present in organisms grown in complex media or isolated from a host and which can interfere with immunologic or biochemical studies. Cesearch on the metabolism of parasites is of immediate interest because pathways that are essential for the parasite but not the host are potential targets for antiprotozoal compounds that would bloc$ that pathway but be safe for humans. Many antiprotozoal drugs were used empirically long before their mechanism of action was $nown. !he sulfa drugs, which bloc$ folate synthesis in malaria parasites, are one example. !he rapid multiplication rate of many parasites increases the chances for mutation# hence, changes in virulence, drug susceptibility, and other characteristics may ta$e place. 4hloro"uine resistance in Plasmodium
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falciparum and arsenic resistance in Trypanosoma rhodesiense are two examples. 4ompetition for nutrients is not usually an important factor in pathogenesis because the amounts utilized by parasitic protozoa are relatively small. Some parasites that inhabit the small intestine can significantly interfere with digestion and absorption and affect the nutritional status of the host# Giardia and Cryptosporidium are examples. !he destruction of the host-s cells and tissues as a result of the parasites- metabolic activities increases the host-s nutritional needs. !his may be a ma;or factor in the outcome of an infection in a malnourished individual. >inally, extracellular or intracellular parasites that destroy cells while feeding can lead to organ dysfunction and serious or life' threatening conse"uences. #.-.#.$C.S 1nglund P!, Sher .eds0A !he %iology of Parasitism. Molecular and Immunological pproach. lan C. (iss, :ew Dor$, &3BB =oldsmith C, 2eyneman / .eds0A !ropical Medicine and Parasitology. ppleton and (ange, 1ast :orwal$, 4!, &3B3 (ee EE, 2utner S2, %ovee 14 .eds0A n Illustrated =uide to the Protozoa. Society of Protozoologists, (awrence, @S, &3B9 @otler /P, Orenstein EMA Prevalence of Intestinal Microsporidiosis in 2I,' infected individuals referred for gastrointestinal evaluation. E =astroenterol B3A &33B, &33+ :eva > , %rown 2A %asic 4linical Parasitology, )th edition, ppleton F (ange, :orwal$, 4!, &33+